加味当归芍药散对淀粉样蛋白42诱导脑内炎症因子及CD45、磷酸化tau蛋白表达的影响

Effects of Modified Danggui Shaoyao Powder on Expression of Cerebral Inflammatory Cytokines, CD45 and Phosphorylated Protein tau Induced by Amyloid Beta 42 in Rats

【目的】观察淀粉样蛋白(Aβ)42沉积对大脑海马炎症因子和CD45、磷酸化tau蛋白(p-tau)表达的影响及中药复方加味当归芍药散的干预作用。【方法】选用SD大鼠,随机分为正常对照组,假手术组,AD模型组,中药高、低剂量组(剂量分别为70.654、35.327 g.kg-1.d-1),西乐葆组(剂量为0.183 g.kg-1.d-1);除前2组外,其他组大鼠均应用立体定位注射方法一次性大脑海马定位注射纤丝状Aβ42复制阿尔茨海默病(AD)病理模型,并给予药物进行干预,采用免疫组织化学方法检测炎症因子及CD45、p-tau表达情况,并用分析软件进行图像分析。【结果】与假手术组比较,AD模型组的白细胞介素-1β(IL-1β)、IL-6及CD45、p-tau的阳性细胞染色面积均显著增加,而染色灰度值均显著下降(P<0.05或P<0.01);与AD模型组比较,各给药组的IL-1β、IL-6阳性染色面积均显著下降(P<0.05或P<0.01);除中药低剂量组IL-1β及西乐葆组IL-6的染色灰度值仅表现为增加的趋势外,其他各给药组的IL-1β、IL-6染色灰度值均显著增加(P<0.05或P<0.01);此外,中药低剂量组和西乐葆组的CD45阳性染色面积均显著下降(P<0.05或P<0.01);各给药组的p-tau染色面积显著下降(均P<0.01);中药低剂量组和西乐葆组的CD45、p-tau染色灰度值均显著升高(P<0.05或P<0.01)。【结论】抑制小胶质细胞的激活及IL-1β、IL-6等炎症因子的产生,进而抑制tau蛋白的过度磷酸化,可能是加味当归芍药散拮抗AD的主要作用机制。

Objective To observe the effects of amyloid beta 42 （Aβ42） deposition on the expression of inflammatory cytokines, CD45 and phosphorylated protein tau （p-tau） in the hippocampus of rats as well asthe intervention of Alzheimer＇s disease （AD） by Modified Danggui Shaoyao Powder （DSP） . Methods SD rats were randomized into normal control group, sham operation group, AD model group, Celebrex group （0.183 g.kg-1.d-1）, and high- and low--dose DSP groups （in the dosages of 70.654, 35.327 g.kg-1.d-1 respectively）. Except for the normal control group and sham operation group, rats in the other groups were injected with single- dose fibrillar Aβ42 in the hippocampus by stereotaxis method to replicate AD rat models. During the modeling, the medication groups were interfered with DSP or Celebrex. The expression of inflammatory cytokines, CD45 and p-tau was observed by SABC immunohistochemical method, and immunohistochemical images were analyzed by image analysis software. Results Compared with the sham operation group, the positive stained area of interleukin-1 beta （IL-1β）, IL-6, CD45 and p-tau in the cells of model group was increased and their grey values were decreased （P〈0.05 or P〈0.01） . Compared with the AD model group, the positively stained area of IL-1β, IL-6 and p-tau in the medication groups , and that of CD45 in low-dose DSP group and Celebrex group was decreased （P〈0.05 or P〈0.01）, and their grey values, except for IL-1β in low-dose DSP group, CD45 and p-tau in high-dose DSP group and IL-6 in Celebrex group, were increased （P〈0.05 or P〈0.01） . Conclusion Inhibiting the activation of microglia and the secretion of inflammatory factors IL-1β and IL-6 , which results into the over-phosphorylation of p-tau, may contribute to the major therapeutic mechanism of DSP for AD.